Key result
Tubulator software accurately estimated overall t-tubule density in synthetic cardiomyocytes (R2 = 0.996), outperforming existing programs particularly in cells with sparse or irregular structures.
Why the study?
Existing analysis tools are less appropriate for irregular structures like dyadic proteins and non-rodent t-tubules, creating a need for robust and versatile tools to characterize these subcellular structures during cardiac disease.
Answers are grounded in this paper and cite their sources.
Tubulator provides a freely accessible, automated tool for detailed 3D analysis of t-tubular and dyadic structures in cardiomyocytes.
During cardiac disease, t-tubules and dyads are remodelled and disrupted within cardiomyocytes, thereby reducing cardiac performance. Given the pathological implications of such dyadic remodelling, robust and versatile tools for characterizing these sub-cellular structures are needed. While analysis programs for continuous and regular structures such as rodent ventricular t-tubules are available, at least in two dimensions, these approaches are less appropriate for assessment of more irregular structures, such as dyadic proteins and non-rodent t-tubules. Here, we demonstrate versatile, easy-to-use software that performs such analyses. This software, called Tubulator , enables automated analysis of t-tubules and dyadic proteins alike, in both tissue sections and isolated myocytes. The program measures densities of subcellular structures and proteins in individual cells, quantifies their distribution into transversely and longitudinally oriented elements, and supports detailed co-localization analyses. Importantly, Tubulator provides tools for three-dimensional assessment and rendering of image stacks, extending examinations from the single plane to the whole-myocyte level. To provide insight into the consequences of dyadic organization for synchrony of Ca 2+ handling, Tubulator also creates ‘distance maps', by calculating the distance from all cytosolic positions to the nearest t-tubule and/or dyad. In conclusion, this freely accessible program provides detailed automated analysis of the three-dimensional nature of dyadic and t-tubular structures. This article is part of the theme issue ‘The cardiomyocyte: new revelations on the interplay between architecture and function in growth, health, and disease’.
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Frisk et al. (2022) studied Cardiomyocyte structural analysis (n=37). Tubulator software vs. TTORG and AutoTT software was evaluated on Correlation between actual and estimated overall t-tubule density (R2 = 0.996). Tubulator software accurately estimated overall t-tubule density in synthetic cardiomyocytes (R2 = 0.996), outperforming existing programs particularly in cells with sparse or irregular structures.
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